[Neural bases of memory and spatial navigation]. / Bases neurales de la mémoire et de la navigation spatiale.

The cognitive map is a concept first introduced by Edward Tolman in 1948 to describe the map of the environment stored in the brain. In this review, after a brief mention of the history of this concept, we explore the contributions of place cells and grid cells to the neural basis of the creation and storage of a spatial map. Finally, we discuss how this map is consolidated and stored in the brain. Questioning and advancing our knowledge of the mechanisms of our memory is essential to improve healthy aging of these systems.

Title: Bases neurales de la mémoire et de la navigation spatiale. Abstract: La carte cognitive est un concept introduit pour la première fois par Edward Tolman en 1948 pour décrire la carte de l'environnement stockée dans le cerveau. Dans cette revue, après une brève évocation de l'histoire de ce concept, nous explorerons les contributions des cellules de lieu et des cellules de grille aux bases neurales de la création et de l'archivage de cette cartographie spatiale. Nous discuterons enfin de la façon dont cette carte est consolidée et stockée dans le cerveau. L'exploration toujours plus poussée des mécanismes de notre mémoire demeure essentielle pour espérer soutenir les adaptations naturelles qui sous-tendent la flexibilité de la cognition au cours de la vie.

Epileptic seizure clustering and accumulation at transition from activity to rest in GAERS rats.

Knowing when seizures occur may help patients and can also provide insight into epileptogenesis mechanisms. We recorded seizures over periods of several days in the Genetic Absence Epileptic Rat from Strasbourg (GAERS) model of absence epilepsy, while we monitored behavioral activity with a combined head accelerometer (ACCEL), neck electromyogram (EMG), and electrooculogram (EOG). The three markers consistently discriminated between states of behavioral activity and rest. Both GAERS and control Wistar rats spent more time in rest (55-66%) than in activity (34-45%), yet GAERS showed prolonged continuous episodes of activity (23 vs. 18 min) and rest (34 vs. 30 min). On average, seizures lasted 13 s and were separated by 3.2 min. Isolated seizures were associated with a decrease in the power of the activity markers from steep for ACCEL to moderate for EMG and weak for EOG, with ACCEL and EMG power changes starting before seizure onset. Seizures tended to occur in bursts, with the probability of seizing significantly increasing around a seizure in a window of ±4 min. Furthermore, the seizure rate was strongly increased for several minutes when transitioning from activity to rest. These results point to mechanisms that control behavioral states as determining factors of seizure occurrence.

Disrupted Co-activation of Interneurons and Hippocampal Network after Focal Kainate Lesion.

GABAergic interneurons are known to control activity balance in physiological conditions and to coordinate hippocampal networks during cognitive tasks. In temporal lobe epilepsy interneuron loss and consecutive network imbalance could favor pathological hypersynchronous epileptic discharges. We tested this hypothesis in mice by in vivo unilateral epileptogenic hippocampal kainate lesion followed by in vitro recording of extracellular potentials and patch-clamp from GFP-expressing interneurons in CA3, in an optimized recording chamber. Slices from lesioned mice displayed, in addition to control synchronous events, larger epileptiform discharges. Despite some ipsi/contralateral and layer variation, interneuron density tended to decrease, average soma size to increase. Their membrane resistance decreased, capacitance increased and contralateral interneuron required higher current intensity to fire action potentials. Examination of synchronous discharges of control and larger amplitudes, revealed that interneurons were biased to fire predominantly with the largest population discharges. Altogether, these observations suggest that the overall effect of reactive cell loss, hypertrophy and reduced contralateral excitability corresponds to interneuron activity tuning to fire with larger population discharges. Such cellular and network mechanisms may contribute to a runaway path toward epilepsy.

Single-fluorophore biosensors based on conformation-sensitive GFP variants.

The ß-strands of GFP form a rigid barrel that protects the chromophore from external influence. Herein, we identified specific mutations in ß-strand 7 that render the chromophore sensitive to interactions of GFP with another protein domain. In the process of converting the FRET-based protein kinase A (PKA) sensor AKAR2 into a single-wavelength PKA sensor containing a GFP and a quencher, we discovered that the quencher was not required and that the sensor response relied on changes in GFP intrinsic fluorescence. The identified mutations in ß-strand 7 render GFP fluorescence intensity and lifetime sensitive to conformational changes of the PKA-sensing domain. In addition, sensors engineered from the GCaMP2 calcium indicator to incorporate a conformation-sensitive GFP (csGFP) exhibited calcium-dependent fluorescence changes. We further demonstrate that single GFP sensors report PKA dynamics in dendritic spines of neurons from brain slices on 2-photon imaging with a high signal-to-baseline ratio and minimal photobleaching. The susceptibility of GFP variants to dynamic interactions with other protein domains provides a new approach to generate single wavelength biosensors for high-resolution imaging.

Mechanisms of excitation of spinal networks by stimulation of the ventral roots.

It has recently been demonstrated that motoneurons in neonatal rodents release an excitatory amino acid, in addition to acetylcholine, from their central terminals onto Renshaw cells. Although the function of this amino acid release is not understood, it may mediate the excitatory actions of motor axon stimulation on spinal motor networks. Stimulation of motor axons in the ventral roots or muscle nerves can activate the locomotor central pattern generator or entrain bursting in the disinhibited cord. Both of these effects persist in the presence of cholinergic antagonists and are abolished or diminished by ionotropic and metabotropic glutamate antagonists. Calcium imaging in the disinhibited cord shows that a ventral root stimulus evokes ventrolateral activity initially, which subsequently propagates to the rest of the cord. This finding suggests that excitatory interneurons excited by motoneuron recurrent collaterals are located in this region. However, motoneurons do not exhibit short latency excitatory potentials in response to ventral root stimulation indicating that the excitatory effects are mediated polysynaptically. We discuss the significance of these findings.

Excitatory actions of ventral root stimulation during network activity generated by the disinhibited neonatal mouse spinal cord.

To further understand the excitatory effects of motoneurons on spinal network function, we investigated the entrainment of disinhibited rhythms by ventral root (VR) stimulation in the neonatal mouse spinal cord. A brief train of stimuli applied to a VR triggered bursting reliably in 31/32 experiments. The same roots that entrained disinhibited bursting could also produce locomotor-like activity with a similar probability when the network was not disinhibited. The ability of VR stimulation to entrain the rhythm persisted in nicotinic and muscarinic cholinergic antagonists but was blocked by the AMPAR antagonist NBQX. Bath application of the type I mGluR1 receptor antagonist CPCCOEt reduced the ability of both dorsal root and VR stimulation to entrain the disinhibited rhythm and abolished the ability of either type of stimulation to evoke locomotor-like activity. Calcium imaging through the lateral aspect of the cord revealed that VR stimulation and spontaneously occurring bursts were accompanied by a wave of activity that originated ventrally and propagated dorsally. Imaging the cut transverse face of L(5) revealed that the earliest VR-evoked optical activity began ventrolaterally. The optical activity accompanying spontaneous bursts could originate ventrolaterally, ventromedially, or throughout the mediolateral extent of the ventral horn or very occasionally dorsally. Collectively, our data indicate that VR stimulation can entrain disinhibited spinal network activity and trigger locomotor-like activity through a mechanism dependent on activation of both ionotropic and metabotropic glutamate receptors. The effects of entrainment appear to be mediated by a ventrolaterally located network that is also active during spontaneously occurring bursts.

Imaging the spatiotemporal organization of neural activity in the developing spinal cord.

In this review, we discuss the use of imaging to visualize the spatiotemporal organization of network activity in the developing spinal cord of the chick embryo and the neonatal mouse. We describe several different methods for loading ion- and voltage-sensitive dyes into spinal neurons and consider the advantages and limitations of each one. We review work in the chick embryo, suggesting that motoneurons play a critical role in the initiation of each cycle of spontaneous network activity and describe how imaging has been used to identify a class of spinal interneuron that appears to be the avian homolog of mammalian Renshaw cells or 1a-inhibitory interneurons. Imaging of locomotor-like activity in the neonatal mouse revealed a wave-like activation of motoneurons during each cycle of discharge. We discuss the significance of this finding and its implications for understanding how locomotor-like activity is coordinated across different segments of the cord. In the last part of the review, we discuss some of the exciting new prospects for the future.

Noncholinergic excitatory actions of motoneurons in the neonatal mammalian spinal cord.

Mammalian spinal motoneurons are considered to be output elements of the spinal cord that generate exclusively cholinergic actions on Renshaw cells, their intraspinal synaptic targets. Here, we show that antidromic stimulation of motor axons evokes depolarizing monosynaptic potentials in Renshaw cells that are depressed, but not abolished, by cholinergic antagonists. This residual potential was abolished by 2-amino-5-phosphonovaleric acid and 6-cyano-7-nitroquinoxaline-2,3-dione. In the presence of cholinergic antagonists, motor axon stimulation triggered locomotor-like activity that was blocked by 2-amino-5-phosphonovaleric acid. Some cholinergic motoneuronal terminals on both Renshaw cells and motoneurons were enriched in glutamate, but none expressed vesicular glutamate transporters. Our results raise the possibility that motoneurons release an excitatory amino acid in addition to acetylcholine and that they may be more directly involved in the genesis of mammalian locomotion than previously believed.

Calcium imaging of network function in the developing spinal cord.

We have used calcium imaging to visualize the spatiotemporal organization of activity generated by in vitro spinal cord preparations of the developing chick embryo and the neonatal mouse. During each episode of spontaneous activity, we found that chick spinal neurons were activated rhythmically and synchronously throughout the transverse extent of the spinal cord. At the onset of a spontaneous episode, optical activity originated in the ventrolateral part of the cord. Back-labeling of spinal interneurons with calcium dyes suggested that this ventrolateral initiation was mediated by activation of a class of interneurons, located dorsomedial to the motor nucleus, that receive direct monosynaptic input from motoneurons. Studies of locomotor-like activity in the anterior lumbar segments of the neonatal mouse cord revealed the existence of a rostrocaudal wave in the oscillatory component of each cycle of rhythmic motoneuron activity. This finding raises the possibility that the activation of mammalian motoneurons during locomotion may share some of the same rostrocaudally organized mechanisms that evolved to control swimming in fishes.

Electroporation loading of calcium-sensitive dyes into the CNS.

Calcium imaging of neural network function has been limited by the extent of tissue labeled or the time taken for labeling. We now describe the use of electroporation-an established technique for transfecting cells with genes-to load neurons with calcium-sensitive dyes in the isolated spinal cord of the neonatal mouse in vitro. The dyes were injected subdurally, intravascularly, or into the central canal. This technique results in rapid and extensive labeling of neurons and their processes at all depths of the spinal cord, over a rostrocaudal extent determined by the position and size of the electrodes. Our results suggest that vascular distribution of the dye is involved in all three types of injections. Electroporation disrupts local reflex and network function only transiently (approximately 1 h), after which time they recover. We describe applications of the method to image activity of neuronal populations and individual neurons during antidromic, reflex, and locomotor-like behaviors. We show that these different motor behaviors are characterized by distinct patterns of activation among the labeled populations of cells.

Spatiotemporal pattern of motoneuron activation in the rostral lumbar and the sacral segments during locomotor-like activity in the neonatal mouse spinal cord.

We used calcium imaging to visualize the spatiotemporal pattern of motoneuron activity during dorsal root-evoked locomotor-like bursting in the lumbosacral spinal cord of the neonatal mouse. Dorsal root stimuli elicited a tonic discharge in motoneurons on which alternating left-right rhythmic discharges were superimposed. Both the tonic and the rhythmic components could be recorded optically from populations of motoneurons labeled with calcium-green dextran. Optical and electrical recordings revealed that rhythmic signals from different parts of the lumbar (L1, L2) and sacral (S1-S3) segments rose, peaked, and decayed in a rostrocaudal sequence. This pattern gave rise to a rostrocaudal "wave" in the activation of motoneurons during each cycle of locomotor-like activity. A similar rostrocaudal delay was observed during episodes of alternation that occurred in the absence of stimulation, suggesting that this delay was not caused by the train of dorsal root stimuli. It is hypothesized that this behavior may simplify the appropriate sequencing of motoneurons during locomotion.

Locomotor-like activity generated by the neonatal mouse spinal cord.

This report describes locomotor-like activity generated by the neonatal mouse spinal cord in vitro. We demonstrate that locomotor-like activity can be produced either spontaneously or by a train of stimuli applied to the dorsal roots or in the presence of bath-applied drugs. Calcium imaging of the motoneuron activity generated by a train of dorsal root stimuli revealed a rostrocaudally propagating component of the optical signal in the anterior lumbar (L1-L3) and in the caudal segments (S1-S4). We hypothesize that this spatio-temporal pattern arises from a rostrocaudal gradient of excitability in the relevant segments. Our experiments suggest that left/right reciprocal inhibition and NMDA-mediated oscillations are not essential mechanisms underlying rhythmogenesis in the neonatal mouse cord. Finally, our data are discussed in the context of other models of locomotion in lower and higher vertebrates.